Cation Adducts Research Articles

Xe-bearing hydrocarbon ions: Observation of Xe.acetylene+[rad] and Xe.benzene+[rad] radical cations and calculations of their ground state structures

This work reports evidence for novel types of Xe-bearing hydrocarbon radical cations. The Xe.acetylene+ radical cation adduct is observed at nearly room temperature using the mass-selected drift cell technique. The irreversible addition of the Xe atom and the lack of back dissociation to HCCH++Xe is consistent with the calculated binding energy of 0.85eV to be contrasted with the metastable nature of the neutral Xe.acetylene adduct. The observed Xe.benzene+ radical cation appears to be a weakly bound complex stabilized mainly by ion-induced dipole interaction consistent with a calculated binding energy in the range of 0.14⿿0.17eV.

Discrimination of Isomeric Carbohydrates as the Electron Transfer Products of Group II Cation Adducts by Ion Mobility Spectrometry and Tandem Mass Spectrometry.

The rapid and unambiguous distinction of isomeric carbohydrate structures persists as a tremendous analytical challenge. This paper reports the first exploitation of carbohydrate/metal ion interactions in concert with gas-phase ion chemistry to improve discrimination of oligosaccharide isomers by both ion mobility spectrometry and tandem mass spectrometry. This is demonstrated for two isomeric pentasaccharides and two isomeric hexasaccharides, each studied in an underivatized form as their calcium ion adducts, barium ion adducts, and gas-phase electron transfer products thereof. With appropriate selection of the charge carrier, transfer of a single electron to the carbohydrate metal ion adducts resulted in isomer-distinguishing shifts in their ion/neutral collision cross sections and the appearance of unique features in their vibrational activation/dissociation spectra. These findings suggest novel and elegant gas-phase strategies for rapid differentiation of isomeric oligosaccharides.

Ion-neutral collisional cross sections of carbohydrate isomers as divalent cation adducts and their electron transfer products.

As the gravity of glycoscience continues to amass, a commensurate demand for rapid, sensitive, and structurally comprehensive glycoanalytical tools has arisen. Among the most elusive but desirable analytical capabilities is the ability to expeditiously and unambiguously detect, distinguish, and resolve carbohydrates that differ only in their constitutional isomerism and/or stereoisomerism. While ion mobility spectrometry (IMS) has proven a highly promising tool for such analyses, the facility of this method to discriminate larger oligosaccharides is still somewhat limited. In an effort to expand the capabilities of IMS to discriminate among carbohydrate isomers, the present investigation was focused on IMS studies of four trisaccharide isomers, four pentasaccharide isomers, and two hexasaccharide isomers, each as group II metal ion adducts and their corresponding gas-phase electron transfer (ET) products. These studies were also evaluated in the context of previously investigated group I metal ion adducts of the same saccharides. The orientationally averaged ion-neutral collisional cross sections (CCSs) of the various carbohydrate/metal ion adducts were found to be dependent on the structures of specific carbohydrate isomers, sensitive to the electronic characteristics of the bound cation, and responsive to the attachment of an additional electron (in the case of divalent metal ion adducts). Overall, these results underscore the utility of metal ions for probing carbohydrate structure in concert with IMS, and the capacity of gas-phase ion chemistry to expand the menu of such probes.

Polysialylated N-Glycans Identified in Human Serum Through Combined Developments in Sample Preparation, Separations, and Electrospray Ionization-Mass Spectrometry

The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans. Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. Anal. Chem.2014, 86, 6268−627624881670), we are able to significantly extend glycan detection sensitivity and provide expanded glycan coverage. We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans.

The effect of cation size (H+, Li+, Na+, and K+) on McLafferty-type rearrangement of even-electron ions in mass spectrometry

Protonation and alkali-metal cation adduction are the most important ionization processes in soft-ionization mass spectrometry. Studies on the fragmentation mechanism of protonated and alkali-metal-cationized compounds in tandem mass spectrometry are essential and helpful for structural analysis. In some cases, it was often observed that a compound attached by different alkali-metal cations (or proton) exhibits similar fragmentation patterns but the relative abundances of product ions are different. This difference was considered to derive from the different electrostatic interactions of alkali-metal cations (or the bonded effect of proton) with the analyte. The alkali-metal cation with a smaller ionic radius shows stronger electrostatic interaction with the molecule because of its higher charge density. In addition, the bonded effect of the proton is stronger than the electrostatic interaction of the alkali-metal cation. In the present study, which used McLafferty-type rearrangements of even-electron ions ([M + Cat]+, Cat = H, Li, Na, K) as model reactions, the effect of cation size in mass spectrometric fragmentation reactions is highlighted. These considerations were also successfully applied to interpret the similar but distinct fragmentation behavior of proton and alkali-metal cation adducts of a synthetic compound (2-(acetamido(phenyl)methyl)-3-oxobutanoate) and a drug (entecavir).

Substitutional Photoluminescence Modulation in Adducts of a Europium Chelate with a Range of Alkali Metal Cations: A Gas-Phase Study

We present gas-phase dispersed photoluminescence spectra of europium(III) 9-hydroxyphenalen-1-one (HPLN) complexes forming adducts with alkali metal ions ([Eu(PLN)3M](+) with M = Li, Na, K, Rb, and Cs) confined in a quadrupole ion trap for study. The mass selected alkali metal cation adducts display a split hypersensitive (5)D0 → (7)F2 Eu(3+) emission band. One of the two emission components shows a linear dependence on the radius of the alkali metal cation whereas the other component displays a quadratic dependence thereon. In addition, the relative intensities of both components invert in the same order. The experimental results are interpreted with the support of density functional calculations and Judd-Ofelt theory, yielding also structural information on the isolated [Eu(PLN)3M](+) chromophores.

Catalytic Effect of Cesium Cation Adduct Formation on the Decarboxylation of Carboxylate Ions in the Gas Phase

The effect of Cs(+) ligation on the decarboxylation of malonic acids (unsubstituted and methyl-, dimethyl-, ethyl-, and phenyl-substituted) in their carboxylate form was studied in the gas phase using tandem mass spectrometry. The study is based on the comparison of the decarboxylation of the bare monoanion (hydrogen malonates) and of the cesium adduct of the cesium salt (Cs(+) [cesium hydrogen malonates]) under collisional activation. Energy-resolved dissociation curves of the negative and positive ions exhibit major differences. Decarboxylation of the cationic adducts of substituted malonic acid salts occurs at significantly lower collisional activation than for the corresponding bare hydrogen malonate anions. The conclusions from these experiments are supported by DFT calculations. The calculated activation parameters (enthalpy and Gibbs energy) confirm that the cesium cation coordination assists the decarboxylation of the carboxylate form.

De novo sequencing of heparan sulfate oligosaccharides by electron-activated dissociation.

Structural characterization of highly sulfated glycosaminoglycans (GAGs) by collisionally activated dissociation (CAD) is challenging because of the extensive sulfate losses mediated by free protons. While removal of the free protons may be achieved through the use of derivatization, metal cation adducts, and/or electrospray supercharging reagents, these steps add complexity to the experimental workflow. It is therefore desirable to develop an analytical approach for GAG sequencing that does not require derivatization or addition of reagents to the electrospray solution. Electron detachment dissociation (EDD) can produce extensive and informative fragmentation for GAGs without the need to remove free protons from the precursor ions. However, EDD is an inefficient process, often requiring consumption of large sample quantities (typically several micrograms), particularly for highly sulfated GAG ions. Here, we report that with improved instrumentation, optimization of the ionization and ion transfer parameters, and enhanced EDD efficiency, it is possible to generate highly informative EDD spectra of highly sulfated GAGs on the liquid chromatography (LC) timescale, with consumption of only a few nanograms of sample. We further show that negative electron transfer dissociation (NETD) is an even more effective fragmentation technique for GAG sequencing, producing fewer sulfate losses while consuming smaller amount of samples. Finally, a simple algorithm was developed for de novo HS sequencing based on their high-resolution tandem mass spectra. These results demonstrate the potential of EDD and NETD as sensitive analytical tools for detailed, high-throughput, de novo structural analyses of highly sulfated GAGs.

Laser Desorption Ionization Mass Spectrometry of Bioactive Substances by Using 2,4,6-Trihydroxyacetophenone on Cation-substituted Zeolite

By using 2,4,6-trihydroxyacetophenone adsorbed to cation-substituted zeolite, laser desorption/ionization mass spectrometry for six kinds of bioactive substances was carried out. The compounds, which were usually difficult to observe by conventional matrix-assisted laser desorption/ionization mass spectrometry, were ionized by cation adduction from the zeolite surface.

Carbonation and Other Super Saturated Gases as Solution Modifiers for Improved Sensitivity in Solvent Assisted Ionization Inlet (SAII) and ESI

Solvent Assisted Ionization Inlet (SAII) produces ions in a heated inlet to a mass spectrometer from aqueous and aqueous/organic solutions with high sensitivity. However, the use of acid modifiers, which typically aids electrospray ionization, generally results in ion suppression in SAII. Here we demonstrate that the use of carbonation and other super-saturated gases as solution modifiers increases analyte ion abundance and reduces metal cation adduction in SAII. Carbonation is also found to enhance electrospray ionization. The mechanistic and practical utility of carbonation in mass spectrometry is addressed.

The effects of cation adduction upon the conformation of three-helix bundle protein domains

The ubiquitin-binding, three-helix bundle domains of the proteins ubiquilin 1 (UQ1) and hHR23A both exhibited remarkably high, but discrete, ammonium ion adduction when electrosprayed from aqueous ammonium acetate. The degree of adduction was highly charge state dependent with, unusually, the lowest charge states (+3 for UQ1 and +4 for hHR23A) showing almost no adducts and the highest charge states (+5 for UQ1 and +6 for hHR23A) exhibiting adduction with two ammonium cations as the most abundant form. As the charge state of protein ions produced by electrospray ionisation (ESI) is related to solvent-accessible surface area we inferred that the ammonium-carrying ions were of a more open conformation than their protonated counterparts. This was confirmed by ESI-travelling wave ion mobility spectrometry-mass spectrometry (TWIMS-MS), which showed that, although the purely protonated ions were compact, their equivalents bearing one or two ammonium adducts exhibited populations of significantly larger collisional cross section (CCS). We postulate that complexation with the ammonium cation may disrupt a key salt bridge(s) in the compact structure. A similar effect is observed with mono-sodium ion adduction, but this is diminished with each additional sodium ion in the complex to produce more compact structures.

Supercharging with Trivalent Metal Ions in Native Mass Spectrometry

Addition of 1.0 mM LaCl(3) to aqueous ammonium acetate solutions containing proteins in their folded native forms can result in a significant increase in the molecular ion charging obtained with electrospray ionization as a result of cation adduction. In combination with m-nitrobenzyl alcohol, molecular ion charge states that are greater than the number of basic sites in the protein can be produced from these native solutions, even for lysozyme, which is conformationally constrained by four intramolecular disulfide bonds. Circular dichroism spectroscopy indicates that the conformation of ubiquitin is not measurably affected with up to 1.0 M LaCl(3), but ion mobility data indicate that the high charge states that are formed when 1.0 mM LaCl(3) is present are more unfolded than the low charge states formed without this reagent. These and other results indicate that the increased charging is a result of La(3+) preferentially adducting onto compact or more native-like conformers during ESI and the gas-phase ions subsequently unfolding as a result of increased Coulomb repulsion. Electron capture dissociation of these high charge-state ions formed from these native solutions results in comparable sequence coverage to that obtained for ions formed from denaturing solutions without supercharging reagents, making this method a potentially powerful tool for obtaining structural information in native mass spectrometry.

Measuring Gas-Phase Basicities Relative to the Lithium Cation by Mass Spectrometry: A Physical Chemistry Experiment

The determination of the gas-phase basicity of organic bases (ligands) toward the lithium cation may be achieved by mass spectrometry, as described for protonic gas-phase basicity by Sunderlin et al. (J. Chem. Educ.2005, 82, 1071–1073). The lithium cation-bound dimers, generated by electrospray ionization, are dissociated by collision-induced dissociation in a tandem mass spectrometer such as triple quadrupole or quadrupole–time-of-flight instruments. An ion trap instrument is not recommended because of solvent adduct formation from Li+ containing fragments. The relative intensities of the ionic fragments are treated along the kinetic method. This experiment is another illustration of the possibilities offered by mass spectrometry for the determination of thermochemical data on gas-phase species. In this experiment, students have the opportunity to identify and manipulate metal–cation adducts which are frequently observed in mass spectrometry. Additional pedagogical interest may arise from discussion of the bonding with Li+, the structural effects on Li+ basicity, and the specific applications of the lithium ion, in particular its role in energy storage.

Use of procaine and procainamide as derivatizing co-matrices for the analysis of oligosaccharides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Analysis of oligosaccharides by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry often yields only alkali metal cation adducts, which results in lower fragmentation yields and difficulty to retrieve sequence information. Derivatization by reductive amination may be used to promote Y-type glycosidic cleavages. However, this involves time-consuming preparations and purifications with sample loss. Here, procaine and procainamide were used directly as co-matrices with 2,5-dihydroxybenzoic acid (DHB). Acidified 10 g/L procaine hydrochloride or procainamide hydrochloride solutions in water/acetonitrile were added to the oligosaccharide solution one minute before preparing our MALDI targets using DHB with the dried-droplet method. This simple protocol resulted in deposits of very fine homogeneous crystals. Positive ion mass spectra, easily acquired in an automated mode, presented a high percentage of oligosaccharides derivatized as Schiff base or glycosylamine notably detected as protonated molecules [M + H](+). The high abundance of procaine or procainamide on the target did not impede the ionization process, improved the signal-to-noise ratio and eliminated the need to search for 'sweet spots'. Fragmentation of the protonated precursor ions of the derivatives largely favored Y-type glycosidic cleavages. This easy and fast sample preparation, involving low toxicity and easily accessible chemicals, allowed the selection of protonated molecules as precursor ions for post-source decay analyses. This opened the possibility of simplifying sequence retrieval in routine oligosaccharide analyses.

Alkali Metal Cation-Induced Destabilization of Gas-Phase Protein–Ligand Complexes: Consequences and Prevention

Electrospray ionization, now a well established technique for studying noncovalent protein-ligand interactions, is prone to production of alkali metal adducts. Here it is shown that this adduction significantly destabilizes the interactions between two model proteins and their ligands and that destabilization correlates with cation size. For both the [FKBP·FK506] and [lysozyme·NAG(n)] systems, dissociation of the metalated complex occurs at markedly lower collision energies than their purely protonated equivalents. Dependency upon size of the metal(+) demonstrates the importance of electrostatic charge density during the dissociation process. Differences in the gas phase basicities (GBapp) of the multiply charged protein ions and proton and sodium affinities of the ligands explain the observed charge partitioning during dissociation of the complexes. Ion mobility-mass spectrometry measurements demonstrate that metal cation adduction does not induce a significant increase in unfolding of the polypeptides, indicating that this is not the principal mechanism responsible for destabilization. Destabilizing effects can be largely reduced by exposing the electrospray to solvent (e.g., acetonitrile) vapor, a method that acts to reduce the amount of adduct formation as well as decrease the charge states of the resulting ions. This approach leads to more accurate determination of apparent K(D)s in the presence of trace alkali metals.

Tandem mass spectrometry studies of protonated and alkali metalated peptoids: Enhanced sequence coverage by metal cation addition

The fragmentation characteristics of five oligo-peptoids were studied under tandem mass spectrometry conditions. The charged peptoids were produced by protonation and alkali metal cation (Li +, Na +, K +, Rb +, and Cs +) addition. The peptoids were ionized by the MALDI process and the resulting ions were fragmented via collision-induced dissociation (CID) experiments. All charged peptoids fragmented predominantly at the amide bonds. Highly abundant and sequence-dependent fragment ions were observed. The fragmentation patterns for the protonated peptoids and the metal cation adducts were strikingly different. All protonated peptoids fragmented by producing predominantly Y-type ions. The bias towards Y-ions was largely due to the greater proton affinity of the secondary amine at the terminal side of the Y-ions. All alkali metalated peptoids fragmented by producing both Y′- and B′-type ions, suggesting a “mobile metal cation” mechanism. For the peptoids with basic side chains, formation of the most abundant ions corresponded to the cleavage of the amide bonds at or near the basic residue. These results suggest that the metal cations are largely coordinated to the side chain of the basic residue. Chelation between the metal cation and the amino groups of the peptoids is an important factor to stabilize the fragment ions. For the peptoid without a basic side chain, the ion intensity was evenly distributed among all medium sized fragment ions. Fragmentations of protonated and alkali metalated peptoids yielded complementary sequential information, which demonstrated the practical utility of using mass spectrometry methods for de novo sequencing of peptoid libraries generated by combinatorial chemistry.

Oligonucleotide Bioanalysis: Sensitivity Versus Specificity

Oligonucleotide Bioanalysis: Sensitivity Versus Specificity

Differentiation of Isomeric HydroxypyridineN-Oxides Using Metal Complexation and Electrospray Ionization Mass Spectrometry

Differentiation between two isomers of hydroxypyridine N-oxide according to the metal cation adducts generated by electrospray ionization (ESI) was investigated for different metal cations, namely Mg(II), Al(III), Ca(II), Sc(III), Fe(III), Co(II), Ni(II), Cu(II), Zn(II), Ga(III), besides the diatomic cation VO(IV). Protonated molecules of the isomeric hydroxypyridine N-oxides as well as the singly/doubly charged adducts formed from neutral or deprotonated ligands and a doubly/triply charged cation were produced in the gas phase using ESI, recording mass spectra with different metal ions for each isomer. While complex formation was successful for 2-hydroxypyridine N-oxide with trivalent ions, in the case of 3-hydroxypyridine N-oxide, only peaks related to the protonated molecule were present. On the other hand, divalent cations formed specific species for each isomer, giving characteristic spectra in every case. Hence, differentiation was possible irrespective of the metal cation utilized. In addition, quantum chemical calculations at the B3LYP/6-31+G(d,p) level of theory were performed in order to gain insight into the different complexation of calcium(II) with the isomers of hydroxypyridine N-oxide. The relative stability in the gas phase of the neutral complexes of calcium made up of two ligands, as well as the singly charged and doubly charged complexes, was investigated. The results of these calculations improved the understanding of the differences observed in the mass spectra obtained for each isomer.

Doped amorphous silica nanoparticles as enhancing agents for surface-assisted time-of-flight mass spectrometry

This article examines the use of doped amorphous silica nanoparticles for surface-assisted laser desorption/ionisation-time of flight-mass spectrometry (SALDI-TOF-MS) of hydrophilic and hydrophobic compounds. A range of particles with surface aliphatic carboxylic, aminophenyl, phenyl or aminopropyl groups have been produced and these have been doped with carbon black, polyaniline or graphite. The effects of surface groups and dopants on the laser desorption/ionisation process were studied. The key factor in effective LDI was the presence of carbon black dopant carrying carboxyphenyl or phenyl residues for positive and negative ion formation. The second key factor was the presence of hydrophilic surface functional groups for hydrophilic amino acid analytes for their detection in positive or negative mode as protonated or de-protonated species respectively whereas hydrophobic surfaces were need for ionisation via cationisation for the hydrophobic analyte squalene. The mechanism for LDI of these particles appears to involve initial adsorption of the analyte onto the surface of the particle, formation of primary ions via adsorption of laser UV irradiation by carboxyphenyl residues attached to the carbon black network which act in an equivalent way to the matrix in matrix-assisted LDI. This is followed by reaction of the primary ions with neighbouring adsorbed analyte molecules. The latter are then released possibly via thermal desorption following proton donation or acceptance from/to via surface residues such carboxylate groups associated with the carbon black within the dopant. Alternatively in the absence of such proton donor/acceptor residues as with hydrophobic particles, the primary ions are released from the particles during desorption and form cation adducts as sodiated and potassiated species in the gas phase above the surface.

Analysis of deprotonated acids with silicon nanoparticle‐assisted laser desorption/ ionization mass spectrometry

Chemically modified silicon nanoparticles were applied for the laser desorption/negative ionization of small acids. A series of substituted sulfonic acids and fatty acids was studied. Compared to desorption ionization on porous silicon (DIOS) and other matrix-less laser desorption/ionization techniques, silicon nanoparticle-assisted laser desorption/ionization (SPALDI) mass spectrometry allows for the analysis of acids in the negative ion mode without the observation of multimers or cation adducts. Using SPALDI, detection limits of many acids reached levels down to 50 pmol/µl. SPALDI of fatty acids with unmodified silicon nanoparticles was compared to SPALDI using the fluoroalkyl silylated silicon powder, with the unmodified particles showing better sensitivity for fatty acids, but with more low-mass background due to impurities and surfactants in the untreated silicon powder. The fatty acids exhibited a size-dependent response in both SPALDI and unmodified SPALDI, showing a signal intensity increase with the chain length of the fatty acids (C12-C18), leveling off at chain lengths of C18-C22. The size effect may be due to the crystallization of long chain fatty acids on the silicon. This hypothesis was further explored and supported by SPALDI of several, similar sized, unsaturated fatty acids with various crystallinities. Fatty acids in milk lipids and tick nymph samples were directly detected and their concentration ratios were determined by SPALDI mass spectrometry without complicated and time-consuming purification and esterification required in the traditional analysis of fatty acids by gas chromatography (GC). These results suggest that SPALDI mass spectrometry has the potential application in fast screening for small acids in crude samples with minimal sample preparation.